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目的:以蚕蛹中的多糖组分为研究对象,优化提取工艺以及研究其抗癌的生物活性。方法:在单因素试验基础上,以微波时间、微波功率、料液比为自变量,蚕蛹多糖收率为响应值,采用中心组合设计方法研究各自变量及其交互作用对多糖收率的影响。CCK-8法检测蚕蛹多糖提取物对SMMC-7721人肝癌细胞生长的抑制情况。Western blot检测细胞凋亡相关蛋白(Bax、Bcl-2和p53)的表达。结果:优化得到的蚕蛹多糖提取条件为:微波时间23.61 min、微波功率516.25 W、料液比1∶28.26(g/mL),多糖收率为4.9%。CCK-8结果分析表明,蚕蛹多糖提取物对SMMC-7721细胞的增殖具有抑制作用,且呈时间和浓度依赖性(P<0.01),其24、48和72 h的半数抑制浓度(IC50)分别为6.606、4.114、2.569 mg/mL。Western blot检测结果表明,随着多糖提取物浓度增大,SMMC-7721细胞Bax、p53蛋白的表达逐渐升高,细胞Bcl-2蛋白的表达逐渐降低。结论:响应面法优化微波提取蚕蛹多糖,工艺简便。蚕蛹多糖提取物具有一定的抗癌活性。
Objective: To study the polysaccharide components of silkworm pupae, optimize the extraction process and study its anti-cancer biological activity. Methods: Based on the single factor experiment, the polysaccharide yield of silkworm pupae was selected as the response time by the microwave time, microwave power and solid-liquid ratio as the independent variables. The center combination design method was used to study the effect of each variable and its interaction on polysaccharide yield. CCK-8 method was used to detect the inhibition of polysaccharides extract of silkworm pupa on the growth of SMMC-7721 human hepatoma cells. Western blot was used to detect the expression of apoptosis-related proteins (Bax, Bcl-2 and p53). Results: The extraction conditions of polysaccharides were as follows: microwave time 23.61 min, microwave power 516.25 W, solid-liquid ratio 1:28.26 (g / mL) and polysaccharide yield 4.9%. CCK-8 results showed that polysaccharides extracted from silkworm pupae could inhibit the proliferation of SMMC-7721 cells in a time and concentration-dependent manner (P <0.01), and the IC50 values at 24, 48 and 72 h 6.606, 4.114, 2.569 mg / mL. The results of Western blot showed that the expression of Bax and p53 protein gradually increased and the expression of Bcl-2 protein gradually decreased in SMMC-7721 cells with the increase of polysaccharide extract concentration. Conclusion: The response surface method is suitable for microwave extraction of silkworm pupa polysaccharides. Silkworm pupa polysaccharide extract has a certain anti-cancer activity.